Cannibalism and predation among larvae of the Anopheles gambiae complex

Among the aquatic developmental stages of the Anopheles gambiae complex (Diptera: Culicidae), both inter- and intra-specific interactions influence the resulting densities of adult mosquito populations. For three members of the complex, An. arabiensis Patton, An. quadriannulatus (Theobald) and An. gambiae Giles sensu stricto, we investigated some aspects of this competition under laboratory conditions. First-instar larvae were consumed by fourth-instar larvae of the same species (cannibalism) and by fourth-instar larvae of other sibling species (predation). Even when larvae were not consumed, the presence of one fourth-instar larva caused a significant reduction in development rate of first-instar larvae. Possible implications of these effects for population dynamics of these malaria vector mosquitoes are discussed.     

Pyrethroid resistance/susceptibility and differential urban/rural distribution of Anopheles arabiensis and An. gambiae s.s. malaria vectors in Nigeria and Ghana

Resistance to pyrethroid insecticides and DDT caused by the kdr gene in the malaria vector Anopheles gambiae Giles s.s. (Diptera: Culicidae) has been reported in several West African countries. To test for pyrethroid resistance in two more countries, we sampled populations of the An. gambiae complex from south-western Ghana and from urban and rural localities in Ogun State, south-west Nigeria. Adult mosquitoes, reared from field-collected larvae, were exposed to the WHO-recommended discriminating dosage of exposure for 1 h to DDT 4%, deltamethrin 0.05% or permethrin 0.75% and mortality was recorded 24 h post-exposure. Susceptibility of An. gambiae s.l. to DDT was 94-100% in Ghana and 72-100% in Nigeria, indicating low levels of DDT resistance. Deltamethrin gave the highest mortality rates: 97-100% in Ghana, 95-100% in Nigeria. Ghanaian samples of An. gambiae s.l. were fully susceptible to permethrin, whereas some resistance to permethrin was detected at 4/5 Nigerian localities (percentage mortalities 75, 82, 88, 90 and 100%), with survivors including both An. arabiensis Patton and An. gambiae s.s. identified by PCR assay. Even so, the mean knockdown time was not significantly different from a susceptible reference strain, indicating absence or low frequency of kdr-type resistance. Such low levels of pyrethroid resistance are unlikely to impair the effectiveness of pyrethroid-impregnated bednets against malaria transmission. Among Nigerian samples of An. gambiae s.l., the majority from two urban localities were identified as An. arabiensis, whereas the majority from rural localities were An. gambiae s.s. These findings are consistent with those of M. Coluzzi et al. (1979). Differences of ecological distribution between molecular forms of An. gambiae s.s. were also found, with rural samples almost exclusively of the S-form, whereas the M-form predominated in urban samples. It is suggested that 'urban island' populations of An. arabiensis and of An. gambiae s.s. M-form in the rainforest belt of West Africa might be appropriate targets for elimination of these malaria vectors by the sterile insect technique.     

Cytogenetics of the Anopheles gambiae complex in Sudan, with special reference to An. arabiensis: relationships with East and West African populations

The species composition of malaria vector mosquitoes belonging to the Anopheles gambiae complex (Diptera: Culicidae) from >40 localities in Sudan, representing most ecological situations, was determined by analysis of ovarian polytene chromosomes. Of 2162 females, 93% were identified as An. arabiensis Patton and 7% were An. gambiae Giles sensu stricto. No hybrids were found between the two species. Anopheles arabiensis occurred in all but two sites, whereas An. gambiae s.s. was effectively limited to the southernmost, more humid localities. For chromosomal paracentric inversions, the degree of polymorphism was low in An. gambiae s.s. (inversions 2La, 2Rb and 2Rd), higher in An. arabiensis (inversions Xe, 2Ra, b, bc, d1, s; 3Ra, d). Anopheles gambiae samples from Sudan were all apparently panmictic, i.e. they did not show restricted gene flow such as observed among West African populations (interpreted as incipient speciation). Chromosomal inversion patterns of An. gambiae in southern Sudan showed characteristics of intergrading Savanna/Forest populations similar to those observed in comparable eco-climatic situations of West Africa. Anopheles arabiensis was polymorphic for inversion systems recorded in West Africa (2Ra, 2Rb, 2Rdl, 3Ra) and for a novel 2Rs polymorphism, overlapping with inversion systems 2Rb and 2Rd1. Samples carrying the 2Rs inversion were mostly from Khashm-el-Girba area in central-eastern Sudan. In the great majority of the samples all polymorphic inversions were found to be in Hardy-Weinberg equilibrium. Sudan populations of An. arabiensis should therefore be considered as generally panmictic. Anopheles arabiensis shows more inversion polymorphism in west than in east African populations. Sudan populations have more evident similarities with those from westwards than those from eastwards of the Great Rift Valley. The possible influence of the Rift on evolution of An. arabiensis is discussed.     

Interspecific variation in diving activity among Anopheles gambiae Giles, An. arabiensis Patton, and An. funestus Giles (Diptera: Culicidae) larvae.

Anopheline larvae generally inhabit the near-surface of aquatic habitats, but they dive and remain at the bottom of these habitats for some time. This study examined forced and voluntary diving behavior and submergence tolerance in the three major African malaria vectors, Anopheles gambiae Giles, An. arabiensis Patton, and An. funestus Giles. The former two species occur sympatrically in temporal and shallow water bodies, while the latter occurs in permanent deeper water bodies. Anopheles funestus was the most tolerant of submergence, but the larvae tended to halt their descent before reaching the bottom by attaching onto a wall. The difference in diving behavior between An. funestus and the two species in the An. gambiae complex may be an adaptation to their contrasting breeding sites, because the former species must spend considerable energy to surface in its typical breeding sites. Both An. gambiae and An. arabiensis reached the bottom and crawled along the substrate, but An. gambiae voluntarily crawled more often than An. arabiensis. The possible importance of asymmetric bottom-feeding between these two sympatric species is discussed.     

Flight tone of field‐collected populations of Anopheles gambiae and An. arabiensis (Diptera: Culicidae)

Abstract.Laboratory colonies of the human malaria vectors Anopheles gambiae Giles and An. arabiensis Patton have distinct flight tones. If flight tone similarly distinguishes natural populations of these sympatric sibling species, it may play a role in reproductive isolation of swarms that are otherwise behaviourally identical. To assess the fidelity of flight tone differences in natural populations, flight tone was measured in the F1 progeny of mosquitoes of both species captured in western Kenya. Flight tone distributions of wild An . gambiae and An. arabiensis were similar to their laboratory conspecifics. However, interspecies comparisons of flight tone of wild mosquitoes revealed significantly different but overlapping distributions for both sexes. Furthermore, when the effect of body size on flight tone was determined, there was a positive correlation between wing length and flight tone for both sexes of An. gambiae and An. arabiensis , suggesting that mosquito size is a significant variable affecting flight tone. Although these findings diminish any practical benefit of flight tone as a diagnostic tool in species identification, its potential role in pre‐mating species recognition needs further investigation.     

Effect of emigration on cannibalism and intraguild predation in aphidophagous ladybirds

Abstract.  1. The incidence and timing of emigration, cannibalism, and intraguild predation of larvae of three aphidophagous ladybirds (Coleoptera: Coccinellidae), Harmonia axyridis Pallas, Coccinella septempunctata brucki Mulsant, and Propylea japonica Mulsant, relative to the presence of prey was determined in the laboratory in single- and mixed-species populations.
2. In single-species populations, 80% of the larvae of C. s. brucki emigrated prior to the extinction of the aphid population and no larvae were lost due to cannibalism; however > 80% of the larvae of the other two species were still present when the aphid became extinct and the losses due to cannibalism for H. axyridis and P. japonica were 25% and 14% respectively. Finally, 28% of the P. japonica larvae completed their development, whereas no larvae of the other two species became adult.
3. In mixed-species populations, mortality of P. japonica attributable to cannibalism or intraguild predation increased greatly to 60%, whereas that of the other two species remained about the same. Consequently, survival of H. axyridis larvae improved and survival of P. japonica worsened; however the survival of C. s. brucki larvae was not affected by the other two species. Early emigration by C. s. brucki larvae may have enabled them to escape intraguild predation by H. axyridis in this system.     

Distribution of a knockdown resistance mutation (L1014S) in Anopheles gambiae s.s. and Anopheles arabiensis in western and southern Kenya

In Kenya, insecticide-treated mosquito nets (ITNs) distributed to pregnant women and children under 5 years old through various programs have resulted in a significant reduction in malaria deaths. All of the World Health Organization-recommended insecticides for mosquito nets are pyrethroids, and vector mosquito resistance to these insecticides is one of the major obstacles to an effective malaria control program. Anopheles gambiae s.s. and Anopheles arabiensis are major malaria vectors that are widely distributed in Kenya. Two point mutations in the voltage-gated sodium channel (L1014F and L1014S) are associated with knockdown resistance (kdr) to DDT and pyrethroids in An. gambiae s.s. While the same point mutations have been reported to be rare in An. arabiensis, some evidence of metabolic resistance has been reported in this species. In order to determine the distribution of the point mutation L1014S in An. gambiae s.s. and An. arabiensis in southern and western Kenya, we collected larvae and screened for the mutation by DNA sequencing. We found high allelic and homozygous frequencies of the L1014S mutation in An. gambiae s.s. The L1014S mutation was also widely distributed in An. arabiensis, although the allelic frequency was lower than in An. gambiae s.s. The same intron sequence (length: 57 base) found in both species indicated that the mutation was introgressed by hybridization. The allelic frequency of L1014S was higher in both species in western regions, demonstrating the strong selection pressure imposed by long-lasting insecticide-treated nets (LLITN)/ITN on the An. gambiae s.s. and An. arabiensis populations in those areas. The present contribution of the L1014S mutation to pyrethroid resistance in An. arabiensis may be negligible. However, the homozygous frequency could increase with continuing selection pressure due to expanded LLITN coverage in the future.     

Comparative fecundity and associated factors for two sibling species of the Anopheles gambiaecomplex occurring sympatrically in The Gambia

Abstract. For two sibling species of mosquitoes belonging to the Anopheles gambiae complex of malaria vectors, the effects of body size (wing length) and bloodmeal size (haematin excretion) on fecundity of wild females were investigated in The Gambia, West Africa. Freshly blood-fed individuals from sympatric populations of An.arabiensis and An.gambiae sensu stricto were sampled by collection at 07.00–09.00 hours from within bednets during July/August 1993, at the beginning of the rainy season. The possible confounding effect of infection with Plasmodium parasites was removed by eliminating infected mosquitoes from the study samples. An.arabiensis females comprised 75% of the An.gambiae sensu law population and were significantly larger (greater mean wing length) than those of An.gambiae s.s. mosquitoes. Mean egg production per female (for the subsequent gonotrophic cycle, excluding pre-gravids) for the two species was not significantly different, though the relationship between wing length and egg production showed An.gambiae s.s. to be more fecund than the An.arabiensis of the same size. Pre-gravid An.gambiae s.s. had consumed significandy smaller bloodmeals than gravid females but the mean wing length of these two gonotrophic categories was not significantly different. In contrast, An.arabiensis pre-gravids were smaller and had consumed smaller bloodmeals than the gravids.     

Studies on Aedes albopictus larval mass-rearing optimization

To set up a sterile male technique program to control Aedes albopictus (Skuse) in areas in northern Italy, a pilot mass-rearing facility is under development. For this purpose, experiments were carried out to find the optimal larval density for the optimization of the rearing parameters, i.e., to obtain the fastest larval development, the highest larval and pupal survival rate, and large-sized pupae. Several different larval densities, from 40 to 2874 larvae per liter, were tested. For densities from 40 to 600 larvae per liter significant size differences were found among pupae obtained under different larval densities. The larvae raised at the lowest density tended to be smaller and to develop most slowly, i.e., longer pupation time. Also, increasing water volume and depth seemed to negatively affect the pupation success. Compared with the other larval densities tested, the larvae reared at a density of 2874 larvae per liter developed slightly faster and showed higher survival rates, indicating this density as appropriate for the development of a mass rearing, at least using the current larval diet.     

Anopheles arabiensis and An. funestus are equally important vectors of malaria in Matola coastal suburb of Maputo, southern Mozambique

Transmission characteristics of malaria were studied in Matola, a coastal suburb of Maputo, the capital City, in southern Mozambique, from November 1994 to April 1996. The local climate alternates between cool dry season (May-October) and hot rainy season (November-April) with mean annual rainfall 650-850 mm. Saltmarsh and freshwater pools provide mosquito breeding sites in Matola. Malaria prevalence reached approximately 60% among people living nearest to the main breeding sites of the vectors. Plasmodium falciparum caused 97% of malaria cases, others being P. malariae and P. ovale. Potential malaria vector mosquitoes (Diptera: Culicidae) collected at Matola during daytime indoor-resting (n = 1021) and on human bait at night (n = 5893) comprised 12% Anopheles coustani Laveran (93% biting outdoors), 46% An. funestus Giles (68% biting indoors) and 42% An. gambiae Giles sensu lato (60% biting outdoors). All 215 specimens of An. gambiae s.l. identified genetically were An. arabiensis Patton. Anopheles funestus populations remained stable throughout the year, whereas densities of the An. gambiae complex fluctuated considerably, with An. arabiensis peaking during the rainy season. No concomitant rise in malaria incidence was observed. Human landing indices of An. funestus and An. arabiensis averaged 1.8 and 3.8 per man-night, respectively. Overall Plasmodium sporozoite rates were 2.42+/-1.24% in 2181 An. funestus and 1.11+/-1.25% in 1689 An. arabiensis dissected and examined microscopically. Mean daily survival rates were 0.79 for both vector species. Estimated infective bites/person/year were 15 An. funestus and 12 An. arabiensis. Biting rates were greatest at 2100-24.00 hours for An. funestus (68% endophagic) and 21.00-03.00 hours for An. arabiensis (40% endophagic). The entomological inoculation rate (EIR) declined sharply over very short distances (50% per 90m) away from breeding-sites of the vectors. Consequently, P. falciparum prevalence among Matola residents was halved 350 m within the town. Implications for the protective effectiveness of a 'cordon sanitaire' by residual house-spraying and/or the use of insecticide-treated bednets are discussed.     

Assessment of Aedes albopictus (Skuse) (Diptera: Culicidae) clutch size in wild and laboratory populations

Aedes albopictus (Skuse) is an invasive mosquito species found across the southern U.S. with range expansion into many northern states. Intra‐ and interspecific larval competition have been evaluated for Ae. albopictus with respect to subsequent adult size, immature and adult survivability, and its capacity to vector pathogens as an adult. However, limited data are available on egg production as related to larval rearing conditions. Because Ae. albopictus is a container‐inhabiting mosquito that oviposits in resource‐limited habitats, it is found under variable density‐dependent conditions. Therefore, we examined the impact of specific rearing conditions on Ae. albopictus clutch size and adult body size; comparing the egg production values and wing lengths from known developmental densities to those from field‐collected populations. Field populations varied significantly among collection sites in mean clutch size (23 to 46). These clutch sizes were comparable to the mean clutch sizes of females reared at the larval densities of nine (20 eggs) and three (53 eggs) larvae per 3 ml of water in the laboratory. Field populations experienced density‐dependent effects impacting adult mosquito size. Mosquitoes from the four sample sites had mean wing lengths of 1.99, 2.47, 2.51, and 2.54 mm, which were less than the mean wing length of mosquitoes reared at larval densities of three larvae per 3 ml of water (2.57 mm).     

The impact of permethrin-impregnated bednets on malaria vectors of the Kenyan coast

Abstract. The effects of introducing permethrin-impregnated bednets on local populations of the malaria vector mosquitoes Anopheles funestus and the An.gambiae complex was monitored during a randomized controlled trial at Kilifi on the Kenyan coast. Pyrethrum spray collections inside 762 households were conducted between May 1994 and April 1995 after the introduction of bednets in half of the study area. All-night human bait collections were performed in two zones (one control and one intervention) for two nights each month during the same period. PCR identifications of An.gambiae sensu lato showed that proportions of sibling species were An.gambiae sensu stricto > An.merus > An.arabiensis.
Indoor-resting densities of An.gambiae s.l. and the proportion of engorged females decreased significantly in intervention zones as compared to control zones. However, the human blood index and Plasmodium falciparum sporozoite rate remained unaffected. Also vector parous rates were unaltered by the intervention, implying that survival rates of malaria vectors were not affected. The human-biting density of An.gambiae s.l. , the predominant vector, was consistently higher in the intervention zone compared to the control zone, but showed 8% reduction compared to pre-intervention biting rates - versus 94% increase in the control zone.
Bioassay, susceptibility and high-performance liquid chromatography results all indicated that the permethrin content applied to the nets was sufficient to maintain high mortality of susceptible vectors throughout the trial. Increased rates of early outdoor-biting, as opposed to indoor-biting later during the night, were behavioural or vector composition changes associated with this intervention, which would require further monitoring during control programmes employing insecticide-treated bednets.     

Comparative responses of ovipositing Anopheles gambiae and Culex quinquefasciatus females to the presence of Culex egg rafts and larvae

Field observations have demonstrated that gravid Anopheles gambiae Giles s.s. (Diptera: Culicidae) are selective in their choice of oviposition sites. For example, immature stages of An. gambiae s.s. are rarely found in water that contains Culex quinquefasciatus Say immatures. The possibility that this may, in part at least, reflect a response by ovipositing An. gambiae s.s. females to volatile signals associated with Culex juveniles was evaluated by testing the response of An. gambiae s.s. females to varying densities of Cx. quinquefasciatus egg rafts and/or larvae in oviposition choice assays. For comparison, the oviposition choices of Cx. quinquefasciatus to conspecific egg rafts and/or larvae were similarly assayed. At a low density of Cx. quinquefasciatus egg rafts (1-15 egg rafts/100 mL water), An. gambiae s.s. females laid more eggs in the treatment water than in the control, with a maximum of twice as many in the treatment water at 5 egg rafts/100 mL water. At higher egg raft densities and in all treatments that included Cx. quinquefasciatus larvae, oviposition decreased significantly in the treatment dishes in a density-dependent manner. As previous studies have indicated, ovipositing Cx. quinquefasciatus females were attracted to and laid egg rafts in dishes containing conspecific egg rafts and, interestingly, also in dishes containing larvae.     

Ovicidal and larvicidal activity against Aedes aegypti and Anopheles gambiae complex mosquitoes of essential oils extracted from three spontaneous plants of Burkina Faso

Essential oils extracted from dried leaves of three spontaneous plants naturally growing in Burkina Faso, i.e. Cymbopogon proximus, Lippia multiflora and Ocimum canum, exhibited larvicidal activity by the WHO standard protocol against 3rd and 4th instar F1-larvae of field-collected mosquitoes vectors of human disease, namely Aedes aegypti and members of the Anopheles gambiae complex, An. arabiensis and An. gambiae. The median lethal concentration (LC50) for Ae. aegypti and An. gambiae s.l. larvae ranged between 53.5-258.5 ppm and 61.9-301.6 ppm, respectively. The LC90 estimates ranged 74.8-334.8 ppm for Ae. aegypti, and 121.6-582.9 ppm for An. gambiae s.l. Ovicidal activity against eggs of An. gambiae s.l. was also demonstrated. The LC50 values for An. gambiae s.l. eggs ranged between 17.1-188.7 ppm, while LC90 values ranged between 33.5-488 ppm. Lippia multiflora showed the highest activity against An. gambiae s.l. eggs and Ae. aegypti larvae, whereas no difference was found among C. proximus and L. multiflora in their activity against An. gambiae s.l. larvae. Of the three plants, essential oils from O. canum had the lowest activity against both eggs and larvae. Eggs were more susceptible than larvae. Ae. aegypti larvae were more susceptible than larvae of An. gambiae s.l.     

Responses of Mamestra brassicae (Lepidoptera: Noctuidae) to crowding: interactions with disease resistance,colour phase and growth

This study examines phenotypic plasticity in relation to rearing density in larvae of the moth, Mamestra brassicae. Larval phase, growth rate, weight at moulting and susceptibility to disease were quantified when reared at five densities. Larvae develop more quickly, but attain a smaller size and are more susceptible to disease, when reared at high than at intermediate densities. They also exhibit a higher degree of melanisation than larvae reared at intermediate densities, or singly. A review of the literature suggests that a switch to a rapidly developing dark phase at high densities is a widespread phenomenon within the Lepidoptera. Rapid development at the expense of attaining a large size, and increased melanisation, are interpreted as adaptive responses to reach pupation before food supplies are depleted, as is likely when larval density is high. High susceptibility to viral infection at high density may be a result of physiological stress associated with rapid development, or due to a shift in allocation of resources from resistance to development: larvae that developed quickly were more susceptible to infection. Larvae reared singly appeared to be less fit than larvae reared at intermediate densities: they exhibited many of the characteristics of larvae reared at high density, particularly low weight, a right-hand skew in their weight frequency distribution, and high susceptibility to disease. I hypothesise that expression of resistance may be phenotypically plastic with regard to environment. Contact with other larvae may, up to a point, stimulate both growth and resistance to infection, for the risk of infection will increase with the density of conspecifics.     

Cannibalism of Helicoverpa zea (Lepidoptera: Noctuidae) from Bacillus thuringiensis (Bt) transgenic corn versus non-Bt corn

Because of the importance of cannibalism in population regulation of Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) in corn, Zea mays L., it is useful to understand the interactions between Bacillus thuringiensis (Bt) transgenic corn and cannibalism. To determine the effects of Bt corn on cannibalism in H. zea, pairs of the same or different instars were taken from Bt or non-Bt corn and placed on artificial diet in proximity. Cannibalism occurred in 91% of pairs and was approximately 7% greater for pairs of larvae reared from Bt transgenic corn (95%) than from non-Bt corn (88%). Also, first instar by first instar pairs had a lower rate of cannibalism than other pairs. Time until cannibalism was not different for larvae from Bt corn versus non-Bt corn. Pupation rate of cannibals and surviving victims was not different for pairs from Bt corn versus non-Bt corn. Finally, cannibalism increased pupation rate of cannibals from both Bt and non-Bt corn by approximately 23 and 12%, respectively, although the increases were not significant. Thus, negative effects of Bt on larvae were compensated by increased cannibalism in comparison with larvae reared on non-Bt corn, which increased larval survival to levels comparable with larvae reared on non-Bt plants.     

Revisiting the role of introgression vs shared ancestral polymorphisms as key processes shaping genetic diversity in the recently separated sibling species of the Anopheles gambiae complex

The role of interspecific hybridisation in the evolution of pest species is poorly understood. In mosquito disease vectors this is of particular importance due to the evolution of insecticide resistance and the proposed release of transgenic strains that are refractory to the malaria parasite. In this study, we apply population genetic methods in a novel manner to determine whether mitochondrial DNA sequences have introgressed between the closely related African malaria vectors Anopheles gambiae and A. arabiensis. Our results suggest that speciation was geologically recent and ancestral haplotypes at the ND5 locus are retained in both species. In addition, comparing haplotype frequencies in allopatric and sympatric populations, suggest locale specific unidirectional introgression of mitochondria from A. arabiensis into A. gambiae.     

Cytogenetic and biometric observations on members of the Anopheles gambiae complex in Mozambique

Four species of the Anopheles gambiae complex were identified in Mozambique (East Africa) by chromosomal analysis. They were An. merus, An. gambiae s.s., An. arabiensis and An. quadriannulatus. An. merus was observed in coastal zones as well as in inner areas where the rivers are tidal and brackish and/or the soil is salty. An. gambiae s.s. is present in the central-northern regions (north of Save river) from the coast to the western mountains. On the coast it is often sympatric with An. merus. It is apparently absent south of Save river. An. arabiensis was observed in samples from the north-western hilly and mountainous areas, sympatrically with An. gambiae s.s., as well as south of Save river where often it is sympatric with An. merus. Only one specimen of An. quadriannulatus was observed. It was from a small sample collected feeding on bovid in a southern locality (Bela Vista-Maputo area). No inversion polymorphism was observed in the 446 An. merus identified. A quite low degree of inversion polymorphism was shown by both An. gambiae s.s. and An. arabiensis, involving 2Rb, 2La and 2Rb inversion systems respectively. By morphological analysis of cytogenetically identified samples of three species of the complex, number of sensilla coeloconica and palpal ratio were confirmed to be useful to distinguish An. merus from An. gambiae s.s./An. arabiensis. The overlapping areas between brackish and freshwater species become smaller when both characters are considered together, 1.5% and 3.5% being the probabilities of confusing An. merus with An. gambiae s.s. and An. arabiensis respectively.     

Observations on the Anopheles gambiae complex in the Senegal River Basin, West Africa

1. Three sibling species of mosquitoes of the Anopheles gambiae complex are found in the Senegal River Basin: An. melas Theobald, An. gambiae Giles and An. arabiensis Patton. 2. An. melas is restricted to the river delta and environs where saltwater breeding places are present. 3. An. gambiae and An. arabiensis are sympatric in the study area; An. arabiensis predominates in coastal zones where it breeds also during the dry season; An. gambiae predominates in inland areas where breeding is mostly restricted to the rainy season (July-September). 4. An. arabiensis is chromosomally polymorphic all over the study area, with much variation of inversion frequencies, particularly for the 2Ra arrangement. 5. An. gambiae is characterized by a very high frequency of the 2Rb-2La inversion arrangement which is typical of the Savanna chromosomal form.     

Distribution of the species of the Anopheles gambiae complex and first evidence of Anopheles merus as a malaria vector in Madagascar

BACKGROUND: Members of the Anopheles gambiae complex are amongst the best malaria vectors in the world, but their vectorial capacities vary between species and populations. A large-scale sampling of An. gambiae sensu lato was carried out in various bioclimatic domains of Madagascar. Local abundance of an unexpected member of this complex raised questions regarding its role in malaria transmission. METHODS: Sampling took place at 38 sites and 2,067 females were collected. Species assessment was performed using a PCR targeting a sequence in the IGS of the rDNA. Analysis focused on the relative prevalence of the species per site, bioclimatic domain and altitude. Infectivity of Anopheles merus was assessed using an ELISA to detect the presence of malarial circumsporozoite protein in the head-thorax. RESULTS: Three species were identified: An. gambiae, Anopheles arabiensis and An. merus. The distribution of each species is mainly a function of bioclimatic domains and, to a lesser extent, altitude. An. arabiensis is present in all bioclimatic domains with highest prevalence in sub-humid, dry and sub-arid domains. An. gambiae has its highest prevalence in the humid domain, is in the minority in dry areas, rare in sub-humid and absent in sub-arid domains. An. merus is restricted to the coastal fringe in the south and west; it was in the majority in one southern village. The majority of sites were sympatric for at least two of the species (21/38) and two sites harboured all three species.The role of An. merus as malaria vector was confirmed in the case of two human-biting females, which were ELISA-positive for Plasmodium falciparum. CONCLUSION: Despite the huge environmental (mainly man-made) changes in Madagascar, the distribution of An. gambiae and An. arabiensis appears unchanged for the past 35 years. The distribution of An. merus is wider than was previously known, and its effectiveness as a malaria vector has been shown for the first time; this species is now on the list of Malagasy malaria vectors.